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学科主题: Biophysics; Biotechnology & Applied Microbiology; Chemistry ; Analytical; Electrochemistry; Nanoscience & Nanotechnology
题名: Oligonucleotide ligation assay-based DNA chip for multiplex detection of single nucleotide polymorphism
作者: Jiao-Yu Deng ; Xian-En Zhang ; Yuan Mang ; Zhi-Ping Zhang ; Ya-Feng Zhou ; Qiang Liu ; Hong-Bing Lu, Zhi-Jun Fu
通讯作者: Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China
关键词: oligonucleotide ligation assay ; DNA chip ; single nucleotide polymorphism detection ; T4 DNA ligase ; pentamer ; enzyme-linked assay ; M. tuberculosis
刊名: BIOSENSORS & BIOELECTRONICS
发表日期: 2004
卷: 19, 期:10, 页:1277-1283
收录类别: SCI
部门归属: Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China; Chinese Acad Sci, Wuhan Inst Hydrobiol, Wuhan 430072, Peoples R China; Wuhan TB Prevent & Cure Inst, Wuhan 430030, Peoples R China
摘要: An oligonucleotide ligation assay-based DNA chip has been developed to detect single nucleotide polymorphism. Synthesized nonamers, complementary to the flanking sequences of the mutation sites in target DNA, were immobilized onto glass slides through disulfide bonds on their 5' terminus. Allele-specific pentamers annealed adjacent to the nonamers on the complementary target DNA, containing 5'-phosphate groups and biotin labeled 3'-ends, were mixed with the target DNA in tube. Ligation reactions between nonamers and pentamers were carried out on chips in the presence of T4 DNA ligase. Ligation products were directly visualized on chips through enzyme-linked assay. The effect of G:T mismatch at different positions of pentamers on the ligation were evaluated. The results showed that any mismatch between pentamer and the target DNA could lead to the decrease of ligation, which can be detected easily. The established approach was further used for multiplex detection of mutations in rpoB gene of rifampin-resistant Mycobacterium tuberculosis clinical isolates. (C) 2003 Elsevier B.V. All rights reserved.
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/9520
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

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Jiao-Yu Deng; Xian-En Zhang; Yuan Mang; Zhi-Ping Zhang; Ya-Feng Zhou; Qiang Liu; Hong-Bing Lu, Zhi-Jun Fu.Oligonucleotide ligation assay-based DNA chip for multiplex detection of single nucleotide polymorphism,BIOSENSORS & BIOELECTRONICS,2004,19(10):1277-1283
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